PT1954258E - Materials and methods for treating influenza viral infections with a cysteamine compound - Google Patents

Description

A MATERIAL AND METHODS FOR TREATING VIRAL INFLUENZA INFLUENZA WITH A COMPOSITE OF CYSTEAMINE "

A virus is a small parasite constituted by nucleic acid (RNA or DNA) included in a protein mantle. Viruses can only replicate by infecting a susceptible host cell and directing host cell machinery to produce more viruses. Adsorption of the virus to cells susceptible thereto, as well as the respective penetration, is mediated by glycoproteins (located in the protein mantle). Most viruses are classified into broad categories with. based on the types of nucleic acid formed during their replication, and the path through which the mRNA is produced. In general, viruses have RNA or DNA genetic material, wherein the nucleic acid may be single chain or double stranded.

Important families of DNA-type viruses (also class viruses and Class II viruses, see Harvey, L. et al., Molecular Cell Biology Quart Cell Biology, WH Freeman & Company, 2000)) adenovi ridae and herpesviridae, poxviridae, papovaviridae, densovirinae, and parvovirinae. Included in the virus families are typically classified as RNA type (also classifiable in 2 ΡΕ1954258

Class III and VI, see Molecular Ce 11 Biology) birna-virldae, reoviridae, astoviridae, arterivirus, caliciviriidae, coronaviridae, flaviviridae, picornaviridae, togaviridae, poliovirus, bornaviridae, filoviridae, paramyxovirianae, pneumovirinae, rhabdoviridae , bunyaviridae, and orthomyxoviridae. Influenza, commonly known as " influenza ", is a contagious disease caused by influenza virus, classified as a member of the family crthomyxoviridae. There are three types of influenza viruses that affect humans: Influenza A, B and C. Influenza A viruses have been isolated from many animal species as well as humans, while influenza B and C viruses mainly affect humans.

Influenza viruses are enveloped viruses containing negative single-stranded RNAs that are segmented and encapsidated. The wrap of the influenza virus. is characterized by the presence of two surface glycoproteins: hemagglutinin and neuraminidase. Influenza A and b virions are pleiomorphic and usually have diameters of 80-120 nm. Influenza C virion has many distinctive properties and is therefore easy to distinguish from the most infrequently related virions A and B.

Influenza viruses attack the respiratory system of humans (ie, the nose, throat, and lungs). For example, an infection with influenza A or B can often cause a highly contagious, acute respiratory illness. Infection with Influenza usually includes the following symptoms: fever, headache, tiredness (which may be extreme), dry cough, sore throat, nasal congestion, and diseases in the body.

It is estimated that millions of people in the

United Kingdom: between 10% and 20% of residents

AND . U. Ations of influenza: children 6 to 23 or older. people who long prázo, people Imonares, or renal immune system of the complications that gripe, pneumonia, i, rhinitis, and sinusitis. : > chronic complaints of 3-month pregnant women get influenza every year. Most of this population recovers, usually after a week or two. In some cases, however, complications arise from an infection with inrruenxa. include if nau peusous with higher risks people of mai; months, women live in an institution of care suffering from chronic, cardiac, diabetes, or debilitated conditions. Some examples may arise from an infection grip; bronchitis, s In addition, health. For example, people with asthma may suffer from asthma attacks while the flu is present, and people with chronic congestive heart disease can make it worse if the condition is triggered by the flu.

In the United States, an average of 36,000 people die each year from influenza, and are admitted to a hospital every year in infection. Fort important in hospitalization average 114,000 people as a result of this, influenza viruses have a very morbidity impact which leads to increases in and visits to / from individuals who provide health care. For example, high hospital admission rates are noted for subjects over 65 years of age and also for children under 5 years of age.

In addition, the spread of the virus * through a population can result in epidemics, whose economic impact can be considerable. High mortality rates were observed due to influenza infections during Virology, (1990)). The epidemic has caused deaths in the United States (the Flu Epidemic of 1957, 1968 and 1977 (Fields, Second Edition, Volume 1, pp. 1075-1152).

Periodically the influenza virus causes a worldwide. For example, the 1918 flu pandemic reportedly reported about 20 million worldwide, and 500,000 deaths in the United States Microbiology, Fourth Edition, University of Texas

Medical Branch in Galveston (1996)). in respiratory droplets during periods of 3

Influenza viruses are transmitted predominantly from one person to another through respiratory droplets (also called droplet dissociation) which are released upon coughing and / or sneezing. The flu virus can remain suspended in the air for hours; but they are sensitive to Ciorior and are rapidly 5 ΡΕ1954258 inactivated at temperatures above 50 ° C. The virus can survive 24-48 hours on hard, non-porous surfaces (eg telephone headsets, computer keyboards, door handles, table surfaces and kitchen shelves, toys); it survives 8 hours on cloth, paper and cloth; and survives for five minutes on the hands (see Muir, P, "Treatment of Influenza, Essential CPE."), include typical methods transmitted between infected mucous membranes and infected respiratory droplets in the air, interpersonally counted, counted with contaminated materials (ie, tissue contaminated by infected discharges from a nose or throat). Transmission of the influenza virus through respiratory droplets may have occurred on the day before the person experiences flu-related symptoms. Adults can continue to transmit the virus to others for another three to seven days after the onset of the first symptoms. Unlike adults, children have the ability to transmit the virus for more than a week. Symptoms are usually present one to four days after the virus enters the body. In some cases, a person may be infected with the influenza virus but not show symptoms. During this period, however, these people may transmit the virus to others, 6 ΡΕ1954258

Few methods are available to prevent an influenza infection and a cure is yet to be developed. Methods of avoiding an influenza infection include vaccination and antiviral medications. Three drugs have been approved in the United States (amantadine, rimantadine, and oseltamivir), which are commercially available for use to treat or prevent the disease caused by influenza virus infection. These compounds nevertheless have a maximum ef fi cacy when used prophylactically, which may enable influenza viruses to rapidly develop resistance thereto. See U.S. Pat. 3,352,312 and 3,152,180. Another 5,935,957 has been disclosed; 3,534,084; compounds of which influenza activity has been described in U.S. Pat. 6,271,373; 5,821,243; 5,684,024; 3,592,934; 3,538,160; 3,496,228; and 3,483,254. topical use of implex and influenza. DE 19,817,511 discloses panthenol for the treatment of Herpes

There is a great need for new therapies for the treatment of viral diseases. While on the one hand much progress has been made in developing a number of therapies for the treatment of bacterial infections, there are few viable therapies for the treatment of viruses. As described above, antiviral drugs and vaccines are primarily methods used in the prevention of influenza infections. Ganciclovir, acyclovir and foscarnet are commonly used for the treatment of herpes virus infections. However, such therapies may have substantial side effects based on their deleterious effects on DNA replication of host cells or on their effect vis-à-vis a limited number of viral infections. Furthermore, as stated above, it is known that viruses develop resistance to the therapies, which causes a progressive decline in their effectiveness.

As far as is known, cysteamine compounds have not previously been described as being useful for the treatment of viral infections.

Brief Summary of the Invention for Infections that are Viral Infected by Distemper: The subject invention provides subject subjects to whom influenza has been diagnosed, as well as to prevent viral infections of influenza.

Therefore, in a first aspect thereof, the present invention provides a cysteamine, or a salt thereof, for use in the treatment of a viral infection with influenza in a subject. Lodish, H. et al., Molecular Cell Biology, Fourth Edition, W. H. Freeman & Company (2000), describes various influenza viruses. 8 ΡΕ1954258

In a second aspect, the present invention provides a cysteamine, or a salt thereof, according to claims 1 to 7 for use in reducing the severity, intensity, or duration of complications associated with a viral infection with influenza. the present invention salt, according to 7, for use in a viral infection with

In a third aspect thereof, it provides cysteamine, or any one of claims 1 to treating influenza-associated symptoms.

In a fourth aspect thereof, the present invention provides a cysteamine, or salt thereof, according to any one of claims 1 to 7 for use in preventing the development of a complication related to a viral infection with influenza.

In one embodiment there is provided an effective infection of a fifth aspect, the present invention provides a composition for use in preventing influenza, comprising an amount of an amine, or a salt thereof, and an anti-viral drug .

In a sixth aspect, the present invention provides a composition comprising an effective amount of a cysteamine, or salt thereof, for use in preventing a viral infection with influenza, and a pharmaceutical material for use in the treatment of symptoms associated with inflammation with influenza. And a drug for administration with influenza.

In a seventh aspect, there is provided a cysteamine, or an antiviral thereof, in the form of a preparation together to treat an infection, in particular, in particular the animals. The subject invention relates to human health as an animal, influenza.

The use of a cysteamine compound to treat and / or to prevent infection with the influenza virus is specifically exemplified herein. Administration of a cysteamine compound to a subject prior to acquiring the influenza virus may help to protect the subject from infection with influenza, or at least ensure that symptoms related to the disease caused by the influenza virus develop to a lesser extent than would be observed in the absence of the cysteamine compound.

In another embodiment, it may be a cysteamine compound to avoid and / or delay the development of influenza-related complications in subjects who are at a high risk of contracting such complications. For example, influenza-related complications such as encephalitis, bronchitis, tracheitis, myositis, rhinitis, sinusitis, asthma, bacterial infections (i.e., bacterial infection with Streptococcus aureus, ΡΕ1954258 10

(eg, Haemophilus influenzae R syndrome, staphylococcal monkey infection), complications: atrial fibrillation, myocarditis, pericarditis, neurological complications (ie, confusion, seizures, psychosis, neuritis, Guillain-Barre syndrome, coma, transverse myelitis, encephalitis, encephalomyelitis), toxic shock syndrome, myositis, myoglobinuria, and renal failure, croup, otitis media, viral infections (ie viral pneumonia), pulmonary fibrosis, bronchiolitis pulmonary aspergillosis, inhosphous and myoglobinemia, carotid failure, early fetal deaths and Lardias in pregnant women, increased peri-natal mortality in pregnant women, In the case of congenital anomalies, at birth, they should be removed from the cysteamine compound. can use symptoms related to a

In another embodiment of the invention, a cysteamine compound for relieving influenza in a subject is diagnosed by influenza. One aim is to use other known agents to treat / prevent the disease causing cysteamine compound either alone or in combination with other influenza viruses (ie, vaccines, antiviral drugs) or to treat (ie, antitussives, mucolytics, and / or expectorants, antipyretics and analgesics, nasal decongestants). 11 ΡΕ1954258

In one embodiment, a cysteamine compound is to be used alone or in conjunction with other known agents that are used to treat / prevent viral infection with influenza. Preferably, a cysteamine compound of the invention is to be used before, during or after exposure of a subject to an influenza virus together with known agents that are used to treat / prevent influenza virus disease (i.e., in vaccines, antiviral drugs) or to treat influenza-related symptoms (i.e., antitussives, mucolytics, and / or expectorants, antipyretics and analgesics, nasal decongestants).

In. In a related embodiment, a cysteamine compound is to be used alone or in conjunction with other known agents that are used to treat and / or prevent avian viral infection (AFV). In accordance with the present invention, the cysteamine compound may be for use by injection or oral administration to treat and / or prevent an ATV infection.

Preferably, a cysteamine compound is to be used alone or together with other known agents useful in the treatment and / or prevention of the various subtypes of avian influenza viruses. More preferably, a cysteamine compound of the invention is intended to be used alone or together with other known agents useful in the treatment and / or prevention of H5N1 AIV. A dosage of at least 0.1 mg / ml of cysteamine hydrochloride, more preferably at least 1 mg / ml of cysteamine hydrochloride, and still more preferably at least 2 mg / ml of cysteamine hydrochloride, may be administered at a subject, to treat and / or avoid any of the above listed AIV subtypes, preferably an H5N1 AIV infection.

In some preferred embodiments, the dosage of cysteamine hydrochloride administered for treatment and / or prevention of an H5N1 AIV infection correlates with the concentration of the virus present in the subject. More preferably, the dosage of cysteamine hydrochloride administered in the treatment and / or prevention of H5N1 AIV infection correlates with an initial concentration of about LD50.

According to the daily dosage of one to one subject prior to the subject invention, the cysteamine compound amount administered viral infection with influenza to protect the subject against viral infection may be 10 mg and 3,000 mg. Preferably, a cysteamine compound is from about 50 mg to about 1 g. In a more preferred embodiment, 200 mg to 90 mg of hydrochloride per day is added per 100 g. and avoid / treat (as with the influenza virus beginning of an influenza A, influenza A, influenza B, and by influenz C, or any of its mutants).

According to the present invention, the amount of the daily dosage of a cysteamine compound administered to a subject since the symptoms associated with an influenza viral infection have been reported is from about 10 mg to about 3,000 mg. Preferably, an α-cysteamine compound is administered. between about 200 mg and 1,500 mg per day. In a more preferred embodiment, about 450 mg to about 900 mg of cysteamine hydrochloride per day is administered to a subject to treat and / or ameliorate the severity of influenza-associated symptoms (such as with avian influenza virus , influenza A, influenza B, and influenza C, or any of their mutants).

According to the present invention, the amount of the daily dosage of a cysteamine compound administered to a subject at risk of contracting complications associated with a viral infection with influenza is between about 10 mg and 3,000 mg. Preferably, a cysteamine compound is administered at from about 200 mg to 1,500 mg per day. In a more preferred embodiment, cysteamine hydrochloride (450 mg and 900 mg) is administered daily to and from a subject to prevent and / or delay the onset of complications associated with an influenza virus infection (such as with avian influenza virus, influenza A, influenza B, and influenza C, any of its mutants). ΡΕ1954258 14

Summary Description of Drawings

THE

Figure shows: isteamine as: constituent of coenzyme A. Figure 2 shows a metabolic pathway of isteamine.

Detailed Description of the Invention The invention in viral infections appreciation provides materials for influenza by treating / ameliorating the symptoms associated with viral infections with influenza; and / or by avoiding / delaying the manifestation of complications associated with viral infections with influenza. The term " symptom (s) " as used herein refers to usual signs or indications that a subject is suffering from a particular disease or condition. For example, symptoms associated with a viral infection with influenza, as used herein, refer to usual signs or indications that a subject is infected with an influenza virus. Flu-related symptoms not included in this document include, but are not limited to, fever, headache, exhaustion / fatigue, muscle aches, sore joints, watery eye irritation, nausea, nausea and / or vomiting, cold sores, chest pains, sneezing and respiratory symptoms (ie, 15 ΡΕ1954258 inflamed mucous membranes, sub-sternal burning, nasal discharge, scratchy / inflamed throat, dry cough, loss of smell).

According to the present invention, the symptoms associated with an infection with the influenza virus can begin between 24 and 48 hours after infection, and may begin suddenly. Often the first indication of a flu is chills or a feeling of cold. Fever is common on some of the early days, and the temperature can rise to between 39 ° C and 39.5 ° C. In many cases, subjects feel sick enough to lie in bed for days; the subjects often feel pains throughout the body, more pronounced in the back and legs. a disease OR in a state (i.e., or in the state of the disease), and can be due to causes 1, 2, 3, 4, 5, 6,

As used herein, the term " complication / ions " refers to processes or pathological events occurring during which it does not form an essential part of the disease or condition. Thus, the term medical / clinical complication which is observed in subjects who have been diagnosed as having an infection with influenza virus.

One complication of an influenza virus infection is that infection with the influenza virus can worsen chronic health problems. For example, complications associated with a viral infection without limitation include encephalitis, bronchitis, tracheitis, myositis, rhinitis, 16 ΡΕ1954258 sinusitis, asthma, bacterial infections (i.e., bacterial infections with Streptococcus aureus, bacterial infections by Haemophilus influenzae (ie, atrial fibrillation, myocarditis, pericarditis), Reye's syndrome, neurological complications (i.e., confusion, seizures, psychosis, neuritis, Guillain-Barre syndrome, coma, myelitis), cardiac complications transient, encephalomyelitis), toxic shock syndrome, myositis, myoglobinuria, and renal failure, croup, otitis media, rabies infections (ie viral pneumonia), pulmonary fibrosis, obliterative bronchiolitis, bronchiectasis, exacerbations of asthma, exacerbations of obstructive pulmonary disease pulmonary abscesses, empyema, pulmonary aspergillosis, myositis and myoglobinemia, cardiac haemorrhage, early and late fetal deaths in pregnant women, increases in perinatal mortality in pregnant women, and congenital abnormalities at birth. " influenza virus " , " viruses " use herein, the family Ortomixovindae, B, and influenza C, as well as influenza viruses considered herein to possess two enzymes

The terms " influenza ", of influenza " or " influenza ", as they relate to a virus, including influenza A, influenza and its mutants. Document viruses include those antigenic glycosylated viruses on their surface: haemagglutinin. The various influenza virus types and influenza viruses which can be treated by the methods of the invention are included in the various types of virus and the viruses of the subtypes H1N1, H1N2, H2N2, H3N2, H3N8, H5N3, H5N3, H5N3, H5N8, H5N9, H7N1, H7N2, H7N3, H7N4, H7N7, H7N7, H9N2, and H1O2, included the following subtypes generally known as " Spanish Flu ", " Asian Flu ", " Hong Kong ", " Avian Flu ", Swine Flu " " Swine Flu ", " Swine Flu of Dogs ". The term " subject " as used herein describes an organism, including humans and mammals, to which a treatment is provided with the compositions according to the present invention. Mammalian species benefiting from the present invention include, but are not limited to, primates, chimpanzees, orangutans, humans, monkeys; and pets (ie, companion animals), such as dogs, cats, mice, rats, guinea pigs, and hamsters. " Management together " and " administered together " as used herein include administering a compound or using a therapeutic method with the composition of the invention (a cysteamine compound) in the treatment of a viral infection with influenza or for the treatment of symptoms and / or complications related to a viral infection with influenza.

For a subject who has been diagnosed with an influenza infection, a cysteamine compound may be administered in conjunction with vaccines, anti-viral, antitussive, mucolytic, and / or expectorant drugs; antipyretics and analgesics; nasal decongestants. By way of example, a compound admixed with a cysteamine compound may be provided, such as in a pharmaceutical composition; or the compound and cysteamine may be provided as separate compounds, such as, for example, separately administered pharmaceutical compositions administered consecutively, simultaneously or at different times. Preferably, when the cysteamine compound and the agent known to treat / prevent influenza infection and / or to treat / prevent influenza-related symptoms and complications are administered separately, they will not be administered at such separate heights throughout the time that there can be no interactions between the cysteamine and the known agent.

In. certain antiviral drugs such as amantadine, rimantadine, ribavirin, idoxuridine, trifluridine, vidarabine, acyclovir, gancyclovir, foscarnet, and the like may be administered together with, but not limited to, a vaccination. , zidovudine, didanosine, zalcitabine, stavudine, famciclovir, oseltamivir, and valaciclovir (the materials and / or methods used to treat a viral infection); or with antitussives, mucolytics, and / or expectorants; antipyretics and analgesics; nasal decongestants (materials used to treat symptoms associated with an infection with influenza). 19 ΡΕ1954258 By way of example, a compound for use with a cysteamine compound of the invention may be provided with the cysteamine compound, such as in the same pharmaceutical composition. Alternatively, the compound and cysteamine may be provided as separate compounds, such as, for example, in separate pharmaceutical compositions administered consecutively, simultaneously, or at different times. Preferably, if the cysteamine compound and the agent known to treat / prevent influenza infections and / or to treat influenza-related symptoms and complications are administered separately, they will not be administered in a manner so alternate in time that the cysteamine compound and the known agent can not interact.

As used herein in this document, a cysteamine reference includes those to a " cysteamine compound " includes the various cysteamine salts, in which the pharmaceutically acceptable salts of a cysteamine compound are obtained. The term " pharmaceutically acceptable salt " as used herein refers to any salt of a cysteamine compound which is pharmaceutically acceptable and does not decrease or strongly inhibit cysteamine. Examples of such inorganic or organic salts or salts, such as the acid, the acid, the arate, the trifluoroacetate, the lactate, the maleate sulfate, the maleate sulfate, , citrate, methanesulfonate, phosphate, nitrate, or chloride.

Thus, in one embodiment of the present invention, the advantages of cysteamine as described herein may be achieved by promoting the endogenous production of cysteamine by natural metabolic processes such as through the action of coenzyme A or as a precursor and / or metabolite of (see Figures 1 and 2). This can be achieved, for example, by the administration of pantothenic acid. ; " effective amount ", as used herein, reflects the amount required for the biological response desired, according to the invention in question, the quantum requirement and a compost of the steam is the amount necessary to treat / a r n f e c tio n 1. by infl u e n za; treat r / rr ^ | Symptom 3 S syndicated with injections v i r a i s p o r i n f i n imatio n; and / or prevent / ameliorate the onset of complications associated with influenza viral infections, in a preferred embodiment, the effective amount of a cysteamine compound is the amount necessary to treat / prevent influenza infection; to treat / ameliorate symptoms associated with an influenza infection; and / or to prevent / ameliorate the appearance of complications in patients at increased risk of contracting complications associated with an influenza infection. The improvement in the symptoms and / or severity of the complications may be a decrease in their severity at 5%, 10%, 15%, 20%, 25% 30, 35%, AC. 45%, 95%, γα g, D₂O 2-₂ and * 5 5 55%, 60%, or 99%. 6%, 75%, 80%, 85%, C \ o.. The present invention is applicable in health speculation if non-human subjects, in particular non-human subjects infected with an influenza virus.

With respect to human subjects, the present invention is especially applicable in the treatment and / or prevention of influenza infections, in particular of avian influenza virus infections. In accordance with the present invention, a cysteamine compound is useful in the treatment and / or prevention of infections by various strains of avian influenza, including viruses of the subtypes H1N1, H1N2, H2N2, H3N2, H3N8, H5N1, H5N2, H5N3 , H5N8, H5M9, H7N1, H7N2, H7N3, H7N4, H7N7, H9N2, and H10N7. In one embodiment of the invention, cysteamine hydrochloride is used to treat and / or prevent infection with the H5N1 avian influenza virus in humans or animals. Cysteamine hydrochloride may be administered alone or together with other agents known to be effective in the treatment and / or prevention of influenza infections.

In a related embodiment, a cysteamine compound (such as cysteamine hydrochloride) is administered alone or together with other known agents that are used to treat and / or prevent an avian influenza (AIV) infection. The cysteamine compound may be administered to a subject by injection or by oral route.

Preferably, a dosage of at least 0.1 mg / ml of cysteamine hydrochloride, more preferably at least 1 mg / ml of cysteamine hydrochloride, and still more preferably at least 2 mg / ml of cysteamine hydrochloride, to a subject, to treat and / or to prevent an H5N1 ATV infection.

In some preferred embodiments, the dosage of cysteamine hydrochloride administered in the treatment and / or prevention of an AIV infection (including subtype H1N1, H1N2, H2N2, H3N2, H3N8, H5N1, H5N2, H5N3, H5N8, H5N9, H7 N1, H7 N2, H7 N3, Η.74, H7 N7, H9 N2, and H10 N7) correlates with. the concentration of virus present in the subject. More preferably, the dosage of cysteamine hydrochloride administered in the treatment and / or prevention of an H5N1 AIV infection correlates with a concentration of about LD50 of the virus present in the subject.

The compositions of the invention may be used in a variety of routes of administration, including, for example, orally administrable forms, such as tablets, capsules or the like, or parenterally, intravenously, intramuscularly, transdermally, buccally, subcutaneously, in suppositories, or by another route. These 23 ΡΕ1954258 compositions are referred to herein in the generic manner as " pharmaceutical compositions ". Typically, they may take the form of dosage units, in particular in physically discrete units suitable as units or for human consumption, each unit containing a predetermined amount of active ingredient calculated to produce the desired therapeutic effect in association with one or more other pharmaceutically acceptable ingredients, i.e., diluent or carrier. the cysteamine cations of the subject invention may be formulated according to known methods to prepare pharmaceutical compositions useful. Various formulations are described which are well known and readily available to those skilled in the art. For example, Remington's Pharmaceutical Science (Martin EW [1995] Easton Pennsylvania,

Mack Pi ub 1 ishing C omp any, 19th edition) d writes fo rmutions that can be used in relation to 3. 1. Ώ V Θ Π Ç 3 0 ΘΓΠ 3.p 1Γ Θ Ç 0 * Include ι- in suitable formulations for parenteral administration, for example, sterile aqueous solutions for injection, which may contain antioxidants, buffers, bacteriostats, and solutes, which render the formulations isotonic with the blood of the intended entity; and aqueous and non-aqueous suspensions, which may include suspending agents and thickening agents. Suspensions may be presented in dosage units or in multi-dose containers, for example in sealed ampoules and in vials, and may be stored in a lyophilized state (19) 19425258 only needing to be conditioned with the sterile liquid carrier, water for injections prior to use. Solutions and suspensions may be prepared for forward administration from sterile powders, granules, tablets, etc.. It is to be understood that in addition to the individually mentioned ingredients above, the formulations of the present invention may include other agents conventional in the art, depending on the type of formulation in question.

Formulations containing a cysteamine compound include those suitable for administration by the oral, rectal, nasal, topical (including buccal and sublingual), vaginal, parenteral (including subcutaneous, intramuscular, intravenous, intradermal, intrathecal and epidural) routes, as well as for administration to the eyes. The formulations may conveniently be presented in. unit dosage forms, and may be prepared by any of the methods well known in the art of pharmacy. These methods include the step of associating the cysteamine compound with the vehicle which constitutes one or more of the accessory ingredients. In general the formulations are prepared by uniformly and infi- nously associating the cysteamine compound with liquid carriers or with solid carriers into very small particles, or both, and then, if necessary, forming the product. In some embodiments, the cysteamine compound may be provided in a formulation for use in a skin patch. Administration of a cysteamine compound according to the present invention may be achieved by any method which is suitable and by techniques prescently or prospectively known to those skilled in the art. In a preferred embodiment, a cysteamine compound is formulated into a patentable oral formulation which is easy to consume, such as a pill, a tablet, a tablet, a gum, a beverage, and the like. The article of consumption is then taken at the time, before, or after, if it undergoes a stressful event and / or when it is necessary to increase activiaa and immunological (i.e., after diagnosis of an influenza infection)

According to the invention, compositions containing as active ingredient an effective amount of cysteamine for use in the treatment of an influenza viral infection may also include one or more non-toxic pharmaceutically acceptable carriers or diluents . Examples of such vehicles include

for use in the invention are ethanol, dimethylsulfoxide, glycerol 1, silica, alumina, starch, sorbitol, inositol, xylitol, D-xylose, mannitol, powdered cellulose, microcrystalline cellulose, ocrystalline, talc, colloidal silica, calcium carbonate, magnesium carbonate, calcium phosphate, calcium silicate Aluminum, aluminum hydroxide, sodium starch phosphate, lecithin, and carriers and diluents equivalents. at

To provide the administration of the desired therapeutic treatment, the compositions of the invention will typically contain from about 0.1% to 95% of the total composition including carrier or diluent. The dosage used may vary based on the age, weight, health status, or the gender of the subject being treated.

In one embodiment, the dosage of cysteamine administered to a patient to have the desired reaction is from about 10 mg to about 3,000 mg per day. The intended reaction may include (1) prevention of infection with influenza virus infections; (2) decreases severity, duration, or intensity of symptoms associated with influenza viral infections; and (3) prevention, delay, or a decrease in severity, duration, or intensity of complications related to influenza viral infections. Preferably, the cysteamine hydrochloride is administered daily at about 50 mg to 1000 mg to achieve the desired reaction. In a more preferred embodiment, the dosage of cysteamine hydrochloride administered to a patient to give a desired reaction is between about 200 mg and 900 mg per day.

The following are examples illustrating procedures for practicing the invention. These examples should not be construed as constituting limitations. All percentages are by weight and all proportions in the solvent mixtures are volumetric, unless otherwise stated. 27 ΡΕ1954258

Example 1 - Treatment of Influenza-Related Symptoms

It treated male-infected symptoms (initially discharged a subject of sex with an influenza virus, demonstrating nasal, fever, exhaustion) associated with an influenza infection, with a nasal decongestant and mucolytic medications that did not require medical prescriptions. These drugs were not effective in treating flu-related symptoms in the first 24 hours.

After non-prescription medications proved to be ineffective, the subject was administered by route. a dose of about 700 mg of cysteamine hydrochloride. Within 24 hours, the symptoms associated with influenza infection had disappeared. The subject felt generally healthy.

Example 2 - Study of Antiviral Activity of Cysteamine against H5N1 Avian Influenza Virus: In vitro and in vivo Studies Using Oseltamivir Phosphate to Control Title

According to one embodiment of the invention, cysteamine demonstrates antiviral activity against the H5N1 avian influenza virus. The object of the present invention is especially advantageous because of its unexpected results 28 ΡΕ1954258 against the avian influenza virus. For example, as described below, cysteamine is especially useful in the treatment of H5N1 avian influenza virus infections, even more so than oseltamivir phosphate (the generic name of which is TAMIFLU), which is an anti-cancer drug against avian influenza virus.

Materials and Methods

Cysteamine (hereinafter referred to as TG 21, including 9 9% cysteamine) was omitted from Omega Bio-Pharma (H.K.), Limited. Embryo eggs of chickens free from specific pathogens (SPF) (Beijing, China) were used in this experiment. The CV strain of the H5N1 avian influenza virus was isolated from infected chickens. Roman chickens were purchased from Hebei, not immunized with a vaccine against the avian influenza virus. TAMIFLU® (Roche (China) Ltd., Shanghai, China) was used as described herein.

Toxicity Assessment of TG21 in Embryonic Chicken Eggs

One gram of TG21 was dissolved in 10 mL (1:10) 0.01 mol / L in PBS (1:10, 10 mg / mL) at pH 7.2, and then diluted into a series of dilutions a 1:10 to 1: 5,120. The diluted drug (test groups) or PBS buffer (control group) was injected into the chorioallantoic chambers of 10 day-old embryonated chicken eggs, 5 eggs ΡΕ1954258 for each dilution. The eggs were shaken and the eggs at 370C were monitored twice daily for 5 weeks to observe the survival of the embryos and to calculate the LD50 value (50% deadly viral dose).

Evaluation of the EID50 of the H5N1 Virus of Avian Influenza in Embryonic Hens Eggs

The virus strain CV strain of avian influenza virus is diluted 1/10 with 0.01 M PBS at pH 7.2, between 1 and 10 -10. 0.2 ml of diluted virus (test group e) or PBS buffer (control group) were inoculated into 10-day-old embryonated chicken eggs color-alannotic strains, 5 eggs for each dilution. The eggs were hatched at 37Â ° C and monitored at 3Â ° C 3 times a day. for 5 days to. the survival of embryos is observed. EID50 values (dose infecting 50% of the eggs) were isolated based on the method of

Reed-Muench.

Evaluation of the Antiviral Effect of TG21 against Avian Influenza in Embryo Hens Eggs. 60 120

One gram of TG21 was dissolved in 10 mL, 0.01 mol / l, pH2.2 and PBS (1: 1, 100 mg / ml), and then serially diluted ½ from 1:10 to 1: 5.120. The diluted TG 21 solution was incubated with the same volume of CV strain of H5N1 avian influenza virus at 10 or 100 times its EID50 value, at room temperature for 30, 60 and 30 ΡΕ 19554258 minutes respectively, 0.2 ml of mixed virus drug solution were incubated in the chorioallantoic chambers of embryonic eggs of SPF chickens at 1-day old age. All embryonated eggs were hatched at 37 ° C and monitored twice daily for 5 days to observe survival of the embryos. The IC 50 value was calculated. In order to control TAMIFLU® anti-viral effect over 100-fold the EID50.

Evaluation of the LD50 of Avian Influenza Virus in Chickens.

The original stock of CV strain of avian influenza virus was diluted 1/10 with 0.01 M PBS pH 7.2, between 10-1 and 10-9, and then used to infect chickens with nasal drops, 10 broilers for dilution. Chickens were monitored twice a day for 7 days to observe survival. The LD50 of avian influenza viruses was calculated for chickens, depending on the survival of the animals.

Evaluation of the Effectiveness of TG21 against Avian Influenza in Chickens.

It was administered to Roman chickens at 4 to 6 weeks of age, TG21 through drinking water, with. dosages of 31 ΡΕ1954258 40, 20, 10 mg TG21 / chicken.day, elevated for three days, then chickens were infected with 2.5, 25, 250 times the EID50 in nasal drops, daily for three days. Animals were treated with the same doses of TG21 for five disLS after contamination.

Chickens were monitored twice a day for 7 days. An untreated negative control was run in parallel. The survival of the animals and the efficacy of the TG21 drug were recorded according to the following formula: Efficacy = (date of death in the control group - death rate in the treatment group) / (death rate in the control group) x 100% .

Results 1. Toxicity 120 hours after inoculation of TG21 into the embryonated eggs, some toxicity was detected at high doses from about 100 mg / ml (1:10 dilution) to 25 mg / ml (1:40). LD50 of TG21 for embryonated chicken eggs was 32.1 mg / mL. No side effects were observed for assays below 12.5 mg / mL (1:80). 2, EID50 of Avian Influenza per H5N1 in Embryo Hens Eggs, and LD50 in Chickens.

When more than 109 times the 32 ΡΕ1954258 initial virus (10 10 concentration) was diluted more than 109 times, the embryos survived. According to the Reed Muench Method, the EID50 of avian influenza by H5N1 in embryonated chicken eggs was calculated as 10 ~ 8,17. When the virus stock was diluted to 10 seconds or less, the virus tested was not fatal. The LD50 of the H5N1 avian influenza virus for chickens was 10æl -1æl / 0.2ml. 3. Antiviral Effect of TG21 on H5N1 Influenza Virus in Embryonic Hens.

Prior to inoculation of the embryonated hen eggs, H5N1 avian influenza viruses were treated with a vaccine and were respectively 15.6, 14.9, and 6.8 mg / ml, the treatment periods of 30 , 60, and 120 minutes the 10-fold dose of the viral EID50 cation value. The IC 50 values were respectively 17.5 16.1 mg / mL, when the virus was treated with TG21 different dilutions of TG21 for 30, 60, and 120 minutes. The IC 50 values of TG21 against the viral H5N1 viruses of avian influenza for 30 and 120 minutes prior to inoculation at doses of 10-fold EID50 (see Table 1 below).

In a TAMIFLU®-treated positive control group, TAMIFLU® IC50 antibodies to H5N1 avian influenza virus. a were 25.1 μg / ml and 1 μg / ml, respectively, for treatment periods of 30 and 120 minutes, prior to inoculation, at a 100-fold viral dose 33 ΡΕ1954258 of the control group), all

the EID50. In a group administering no J negative (not if the embryonated chicken eggs died.) TABLE 1 - Antiviral effect of TG21 on chicken eggs embryonated with AIV H5N1 Dl: 10c 100% (16 / 16d) 100% (17/17) 100% (21/21) 100% (16/16) 100% (16/16) 1 1:20 100% (11/11) 100% (12/12) 100% (16/16) 100% (11 / 11) 100% (11/11) 100% (6/6) 100% (6/6) 100% (11/11) ) ç 1:80 37.5% (3/8) 40% (4/10) 66.7% (6/9) 0% (0/5) 20% (1/5) and 1: 160 10% (1/10) 27% (3/11) 36.4% (4/11) 0% (0/10) 0% (0/9) s 1: 320 0% (0/14) 0% (0 / 13) 21% (3/14) 0% (0/15) 0% (0/14) T 1: 640 0% (0/19) 0% (0/18) 13% (2/16) 0 % (0/20) 0% (0/19) G 2 1: 1.280 0% (0/24) 0% (0/23) 0% (0/19) 0% (0/25) 0% (0 / 24) 1 1: 2.560 0% (0/29) 0% (0/28) 0% (0/24) 0% (0/30) 0% (0/29) IC50e IC50 (mg / mL) 15 , 6 14.9 6.8 17.5 16.1 10 EID50â 100 EID50 30 Minutesb 60 Minutes 120 Minutes 30 Minutes 120 Minutes Embryo survival (%)

Note: Mins = Minutes; aEID50: drug dose for 50% infection of eggs; pre-infection drug-virus reaction period; the drug initiation concentration is 100 mg / ml; gross / total; eIC50: drug concentration required for 50% of surviving embryos.

Efficiency of TG21 against H5N1 Avian Influenza Virus in Chickens.

It was administered to broilers at four to six weeks of age, 10-40 mg / f range. day of TG21 in drinking water for three days before and five days after infection with a high dose of virus (250 X LD50), a 34 ΡΕ1954258 mean dose of viras (25 X LD50), and a small dosage of virus (2.5 X LD50) using the H5N1 avian influenza virus. The results of the antiviral effect of the TG21 ranges on the H5N1 avian influenza virus is shown in Table 2 below. All chickens tested died within the first three days after dosages of 250-fold viral infectious LD50, including animals from the control group to TAMIFLU®. This may be due to a dosage of too high viral infection, in such a way that no medicinal treatment, including TAMIFLU, can provide effective protection against viral infection. The protection of TG21 against H5N1 avian influenza virus in broilers at doses of 40, 20, 10 mg / chicken.day -1 were respectively 100%, 62.5%, and 87.5%, respectively, with doses of infection of 2.5 times the infectious dose LD50, and respectively 70%, 80%, and 50%, with 25-fold infectious doses of the LD 50 infectious dose. The statistical difference of the efficiency of TG21 versus the negative control (no drug) was extremely significant (all p <0.01 by the square chi). The efficiency of TAMIFLU® (at 5 mg / chicken.day-1) was 50% for an infection with 25 times the viral dose LD50. No significant difference was found between the IC 50 of TG21 (at 10 mg / chicken .dia-1) and TAMIFLU® (at 5 mg / chicken.day-J) (P &gt; 0.005) for a 25-fold dose infection (Table 2). 35 ΡΕ1954258 TABLE 2 - Antiviral effect of TG21 against H5N1 avian influenza virus in broilers Dose of viral infection Drug and dose 250 LD50 25 LD50 2.5 LD50 TG 40 mg / mL 0% (0 / 10a) 70% (7 / 10) ** 100% (10/10) ** 2 20 mg / mL 10% (0/10) 80% (8/10) ** 62.5% (7/10) ** 10 mg / mL Tamiflu (5.4 mg / mL) 0% (0/10) 50% (5/10) ** 87.5% (9/10) ** N / A Negative Control 0% (0/10) 0% (0/10) 20% (2/10)

Note: ** P &lt; 0.01 compared to the control group by the square χ test; â: Survivors / total

summary

Ten-day embryonated eggs of SPF chickens (free of specific pathogens) and chickens four to six weeks old were used in this study to test the antiviral effect of TG21 on the CV strain of the H5N1 avian influenza virus. The LD50 value (Lice Dose at 50%) of the virus for the chickens, the EID50 (infectious dose of 50% of the eggs) of the virus for embryonated eggs, and the LD 50 of the embryonic eggs treated with TG21 were initially determined. For in vitro studies, viruses were preincubated with different concentrations of TG 21 for 30-120 minutes, respectively, and then the mixture was inoculated into embryonated eggs to observe survival of the embryo. The value of IC 50 (drug concentration required to survive 50% of the embryos) was calculated for the chickens drinking water of TG21. N0S studies in vivo, high, medium and small doses of TG21 were administered for three days before and five days later 36 ΡΕ1954258 from a virus infection. The survival of the animals was recorded and the efficiency of the drug evaluated. Positive control (TAMIFLU®) and negative control (no drug) assays were conducted in parallel studies. The results showed that (1) the IC50 values of TG21 against H5N1 avian influenza viruses in embryonated chicken eggs were 15.6, 14.9 and 6.8 mg / ml, respectively, when 10 times the amount EID50 virus with TG2 1 for 30, 60, 120 minutes &lt; of 17, 5 and 16, 1 mg / ml, respectively 100 times the KID50 virus prevalence of pre-inoculation

cL: D50 virus with TG21 for 30 and 120 minutes prior to inoculation. The IC50U values of TAMIFLU against the H5N1 avian influenza virus in embryonated chicken eggs were 25.1 and 19.4 mg / ml, respectively, when incubating 100-fold the virus EID50 with TAMIFLU for 30 and for 60 minutes prior to inoculation; and (2) the efficiency of TG21 at doses of 40, 20, 10 mg / chicken.day-1 against induced mortality in chickens by the H5N1 avian influenza virus was 100%, 62.5%, and 87 , 5% in the case of a viral infection dose of 2.5 times the dose of LD50 infection and 70%, 80% and 50% after infection with 25 times the viral dose LD50, while the efficiency of TAMIFLU® ( at 5 mg / chicken.day -1), was 50%. These results suggest that cysteamine has a strong antiviral activity against the avian influenza H5N1 virus and may provide similar or even greater protection against the avian influenza H5N1 virus than the commonly licensed drugs against.

avian influenza, such as TAMILFLU 37 ΡΕ1954258

Example 3 - Antiviral activity of Cysteamine against H5N1 avian influenza virus in mice

Materials and Methods

Cysteamine (hereinafter referred to as &quot; TG21 &quot;; including 99% cysteamine) was obtained from Omega Bio-Pharma (H.K.), Limited. The WV strain of the H5N1 avian influenza virus was obtained by isolating it from infected chickens. TAMIFLU * (Roche (China) Ltd., Shanghai, China) was used as described herein.

Evaluation of the dose Lethal to 50% of the H5N1 avian influenza virus in mice (itiLDSO)

A stock solution of the WV strain of the H5N1 avian influenza virus was initially diluted 1: 5, and then diluted with PBS in a series of 5 dilutions H (1: 5 to 1: 1.280). Six to eight week old female mice were anesthetized by intramuscular injection of 100 μl of a 1% sodium barbiturate solution, and then inoculated by dripping 50 μl of diluted WV strain of the H5N1 virus from avian influenza in the nasal cavity of each (n = 10 mice for one of the dilutions). The animals were monitored daily for 14 days and mLD50 was calculated based on the death of the mice using the Reed-Muench method. The results indicate that mouse survival was 0% in the 1: 5, 10% virus dilution group in the 1:20 virus dilution 38 ΡΕ1954258, 25% in the 1:80 virus dilution group , 80% in the 1: 320 virus dilution group, and 90% in the 1: 1280 virus dilution group. The inLD50 value for the WV strain of the avian influenza H5N1 virus was 10Âμl, 13Â ° / 0.05mL or a dilution of 1: 141.5 / 0.05mL.

Therapeutic role of cysteamine in mice infected with avian influenza virus

Fifty female mice (6-8 weeks old) were distributed through three treatment groups (ΤΙ, T2, and T3), one negative (untreated) control group, and one positive control group (TAMIFLU ®) , with 10 mice in each group. After being anesthetized by intramuscular injection of 100 μl of 1% sodium barbiturate, all mice were inoculated intranasally with 10 times the mlD50 of the avian influenza H5N1 virus in 50 μl of PBS. During the first hour after infection, animals were treated by continuing for 12 days by oral dosing with TG21 at a daily dose of 4.8, 2.4, 1.2 mg per mouse respectively , in the T1-T3 treatment groups, daily administration of 0.3 mg TAMILFLU® per mouse in the positive control group and the same volume of PBS in the control group.

The mice were monitored twice daily for 14 days to find clinical signs of infection and to see if they survived. The rate of 39 ΡΕ1954258 av ían, and the groups at equation = (group date at X 100 '0 0, respectively, were calculated with TG21 (protection of TG21 against influenza H5N1 virus compared to For example, to identify the protection rate was (%) death in the control group-death rate in the treatment) / (death rate in the control group) The results showed that the protection was 50%, 70% and 10% in the treatment of the T1 group and 4.8 mg / day in the treatment group T2 (at 2.4 mg / mouse, day 1), and in the treatment of the T3 group (at 1.2 mg / mouse, dial &quot;1); with a protection ratio of 0% η the negative control group and a protection rate of c ns in the positive control group (the TAMIFLU®). The protection rate of TG21 in the Tl group (P &lt; 0.05) in the (P &lt; 0.05) group differed significantly from the rates observed in the (P &lt; 0.01) SJ and J-L / ye group in the T3 group (P &lt; negative control group. These results indicate that TG21 has. a strong antiviral activity against the H5N1 avian influenza virus, and is the ideal drug for the treatment of viral infections due to avian influenza.

Lisbon, October 11, 2012

Claims (17)

A salt thereof, for viral infection by a cysteamine, or use in the treatment of an influenza in a subject. A cysteamine, or a salt thereof, according to claim 1, wherein the viral infection is selected from the group consisting of avian influenza virus and influenza A, B, and C. A cysteamine, or salt thereof, according to claim 2, wherein the subject is infected with at least one avian influenza subtype selected from the group consisting of H1N1, H1N2, H2N2, H3N2, H3N8, H5N1, H5N2, H5N3, H5N8, H5N9, H7N1, H7N2, H7N3, H7N4, H7N7, H9N2, and H10N7. A cysteamine, or salt thereof, according to claim 1, which comprises administering at least 0.1 mg of cysteamine, or a salt thereof, to the subject daily. 5. claim subject between salt. A cysteamine, or a salt thereof, of 4, which comprises daily administration to 2 mg and 3,000 mg of cysteamine, or a pharmaceutically acceptable salt thereof. 6. A process for the preparation of a pharmaceutical composition according to claim 1, wherein said cysteamine salt is cysteamine or cysteamine phosphate. A cysteamine, or a salt thereof, wherein said cysteamine or salt thereof is administered orally, parenterally, intravenously, intramuscularly, transdermally, via the oral route, subcutaneously, or through suppositories. A cysteamine, or a salt thereof, according to any one of claims 1 to 7, for use in reducing the severity, intensity, or duration of the associated cations. viral infection by A cysteamine, or a salt thereof, according to any one of claims 1 to 7, for use in the treatment of symptoms associated with a viral infection by influenza. related A cysteamine, or a salt thereof, any one of claims 1 to 7 for the prevention of the development of a cysteamine. complication with a viral infection by influenza. A viral infection composition for use to prevent influenza, including an effective amount of a cysteamine, that of an SclI, and a drug 3 ΡΕ1954258 The composition according to claim 11, wherein the influenza virus infection is derived from an avian influenza virus and wherein the antiviral drug is selected from the group consisting of ganciclovir, acyclovir, foscarnet, amantadine, rimantadine and oseltamivir. 13. An effective coupling of a cysteamine, or of preventing a viral infection for use in combination with an infection containing an amount of its salt, for use by influenza and a material for the treatment of influenza symptoms. in The composition according to claim 13, wherein the influenza virus infection is derived from an influenza virus, and the virus is infected with influenza virus. .1 tuss d.COS f avian and in. that the pharmaceutical material is comprised of a plurality of pharmaceutically acceptable excipients. analgesics and decongestants; A cysteamine, or a salt thereof, and an antiviral drug in the form of a preparation for use in the treatment of a viral infection by influenza, The cysteamine, or a salt thereof, and the antiviral drug according to claim 15, wherein the influenza virus infection is selected from those derived from the viruses defined in any one of claims 2 or 3. 4 ΡΕ1954258 The cysteamine, or a salt thereof, and the antiviral drug of claim 15 or claim 16, also including any of the features of claims 4 to 7.